Engel J N, Gunning P W, Kedes L
Proc Natl Acad Sci U S A. 1981 Aug;78(8):4674-8. doi: 10.1073/pnas.78.8.4674.
We have utilized cloned actin genes from Drosophila melanogaster and from chicken to isolate 12 actin gene fragments from a human DNA library. Each of these 12 clones was shown to contain actin coding regions by its ability to selectively hybridize to human actin mRNA as assayed by in vitro translation. The translation product was judged to be actin on the basis of its comigration with authentic actins when electrophoresed on one- and two-dimensional NaDodSO4/polyacrylamide gels and on the basis of its partial proteolysis products. Determination of the sizes and order of the fragments generated by restriction endonuclease digestion of each of these recombinant phages allows us to conclude that they are nonallelic and are from nonoverlapping regions of the genome. We have used these cloned human actin genes and the Drosophila and chicken actin gene clones to show that the human genome contains 25-30 EcoRI fragments homologous to actin genes and that, among three nonconsanguineous individuals tested, none of these fragments exhibit length or restriction-site polymorphism.
我们利用从黑腹果蝇和鸡中克隆的肌动蛋白基因，从人类DNA文库中分离出12个肌动蛋白基因片段。通过体外翻译检测，这12个克隆中的每一个都能与人类肌动蛋白mRNA选择性杂交，从而表明它们都含有肌动蛋白编码区。当在一维和二维十二烷基硫酸钠/聚丙烯酰胺凝胶上进行电泳时，根据其与真实肌动蛋白的共迁移情况以及其部分蛋白水解产物，判断翻译产物为肌动蛋白。通过对这些重组噬菌体进行限制性内切酶消化所产生片段的大小和顺序的测定，我们可以得出结论，它们是非等位基因的，且来自基因组的非重叠区域。我们利用这些克隆的人类肌动蛋白基因以及果蝇和鸡的肌动蛋白基因克隆，证明人类基因组包含25 - 30个与肌动蛋白基因同源的EcoRI片段，并且在测试的三个非近亲个体中，这些片段均未表现出长度或限制性酶切位点多态性。
